摘要:
作为重要的本土模式鱼类,稀有鮈鲫神经发育相关的生物学背景几乎空白,导致其在神经毒性评价方面的应用受到限制。本文从稀有鮈鲫脑部克隆得到了神经发育相关的神经生长因子(nerve growth factor;ngf)、脑源性神经营养因子(brain-derived neurotrophic factor;bdnf)、胶质纤维酸性蛋白(glial fibrillary acidic protein;gfap)、髓鞘相关糖蛋白(myelin-associated glycoprotein;mag)、P0蛋白(myelin protein zero;mpz)、微管蛋白α1(α1-tubulin)和Y染色体性别决定基因10(SRY-box containing gene 10;sox10)等基因的片段,并对其进化树及表达谱进行了分析。序列分析表明,bdnf的核苷酸序列与鲫鱼、鲤鱼的同源性最高,均为98%;gfap、ngf、mag和α1-tubulin与斑马鱼的同源性最高,分别为95%、92%、92%和96%;mpz与黑头软口鲦的同源性最高,为94%;sox10与鳙鱼同源性最高,达97%。表达谱分析结果表明,ngf、bdnf、mag、mpz、α1-tubulin和sox10基因均在脑组织中表达量最高,gfap在脊髓中表达量最高,上述结果为这一模式鱼类在神经发育学和神经毒性效应评价方面的应用奠定了基础。
Abstract:
The poverty in knowledge of neurodevelopment of Chinese rare minnow (Gobiocypris rarus) limited its application as a native model fish in neurotoxicital studies and risk assessment. In this case, partial of nerve growth factor (ngf), brain-derived neurotrophic factor (bdnf), glial fibrillary acidic protein (gfap), myelin-associated glycoprotein (mag), myelin protein zero (mpz), α1-tubulin and SRY-box containing gene 10 (sox10) were cloned from brain of rare minnow, and their sequences and transcriptional profile were also studied. Blasting results demonstrated that the sequences of gfap, ngf, mag andα1-tubulin in rare minnow shared the highest identities with those of zebrafish (Danio rerio) (95%,92%,92% and 96%, respectively). Bdnf gene shared the highest homology with the nucleotide sequence of Carassius auratus and Cyprinus carpio (98%), whereas mpz and sox10 were more homogenous with the nucleotide sequence of Pimephales promelas (94%) and Hypophthalmichthys nobilis (97%) respectively. Besides, the transcriptional profile revealed highest abundance of ngf, bdnf, mag, mpz, α1-tubulin and sox10 mRNA in the brain and of gfap in the spinal cord of Chinese rare minnow. These results provided a foundation for future studies in neurodevelopment and neurotoxicity assessment using rare minnow as a native model fish.